产品说明
一般描述
辣根过氧化物酶是从辣根(Amoracia rusticana)中分离出来的,属于过氧化物酶的铁原卟啉基团。HRP是含有四个二硫键的单链多肽。它是一种含有18%碳水化合物的糖蛋白。碳水化合物由半乳糖、阿拉伯糖、木糖、岩藻糖、甘露糖、甘露糖胺和半乳糖胺组成,具体取决于特定同工酶。其分子量(约44 kDa)包括多肽链(33,890道尔顿)、氯化血红素加Ca2+ (约700道尔顿)和碳水化合物(约9,400道尔顿)。至少存在7种HRP同工酶。 辣根过氧化物酶同工酶的等电点范围为3.0-9.0。
应用
The enzyme from Sigma has been used while assessing the skin sensitization potential of pro-haptens. It has also been used to show that peroxidase (PO) activity and its heat stability correlate with the availability of free Ca2+ ions.
Horseradish peroxidase (HRP) is isolated from horseradish roots (Amoracia rusticana). It is used in biochemistry applications such as western blots, ELISA and Immunohistochemistry. Horseradish peroxidase is used to amplify a weak signal and increase detectability of a target molecule, such as a protein. Product P6140 has been used to detect low density lipoprotein (LDL).
生化/生理作用
HRP readily combines with hydrogen peroxide (H2O2) and the resultant [HRP-H2O2] complex can oxidize a wide variety of hydrogen donors. The optimal pH is 6.0-6.5 and the enzyme is most stable in the pH range of 5.0-9.0. HRP can be conjugated to antibodies by several different methods that include the use of glutaraldehyde, periodate oxidation, disulfide bonds, and also via amino- and thiol-directed cross-linkers. It is smaller and more stable than the enzyme labels, β-galactosidase and alkaline phosphatase. Hence, it is the most desired label. Also, its glycosylation leads to lower non-specific binding. It is also used for the determination of glucose and peroxides in solution. Sodium azide, cyanide, L-cystine, dichromate, ethylenethiourea, hydroxylamine, sulfide, vanadate, p-aminobenzoic acid, and Cd2+, Co2+, Cu2+, Fe3+, Mn2+, Ni2+, Pb2+ ions are found to inhibit its enzyme activity.
When incubated with a substrate, horseradish peroxidase produces a coloured, fluorimetric, or luminescent derivative of the labeled molecule, allowing quantification. Horseradish peroxidase has been shown to slightly reduce the level of inhibition in a cydAB mutant.
单位定义
一个连苯三酚单元将在20℃、pH 6.0条件下,在20秒内从连苯三酚形成1.0mg的红棓酚。
外形
Crystalline suspension in 3.2 M (NH4)2SO4 solution containing potassium phosphate buffer, pH 6.0
制备说明
Water may be used to dilute suspension if needed.
分析说明
RZ(Reinheitszahl)是指溶于去离子水中的酶(0.5–1.0 mg/ml)的吸光度比值A403/A275 。 它是血红素含量的量度,而不是酶活性。 即使是RZ值较高的制剂也可能具有较低的酶活性。
Preliminary studies indicate the presence of two basic and no acidic isoenzymes
其他说明
有关 过氧化物酶 的更多信息,请访问 www.sigma-aldrich.com/enzymeexplorer。
基本信息
酶学委员会(EC)编号 | 1.11.1.7 ( BRENDA | IUBMB ) |
EC 号 | 232-668-6 |
MDL编号 | MFCD00071339 |
eCl@ss | 32160410 |
NACRES | NA.54 |
产品性质
质量水平 | 200 |
类型 | Type X |
形式 | ammonium sulfate suspension |
specific activity | ≥225 units/mg protein (biuret, using pyrogallol) |
分子量 | ~44 kDa |
浓度 | ≥1.0 mg/mL |
吸光度比值 | RZ 2.5-3.5 |
储存温度 | 2-8℃ |
安全信息
象形图 | |
警示用语: | Danger |
危险声明 | H317 - H334 |
预防措施声明 | P261 - P272 - P280 - P284 - P302 + P352 - P333 + P313 |
危险分类 | Resp. Sens. 1 - Skin Sens. 1 |
储存分类代码 | 11 - Combustible Solids |
WGK | WGK 3 |
个人防护装备 | Eyeshields, Gloves, type N95 (US) |